Introduction to NGS. Josef K Vogt Slides by: Simon Rasmussen Next Generation Sequencing Analysis
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1 Introduction to NGS Josef K Vogt Slides by: Simon Rasmussen 2017
2 Life science data deluge Massive unstructured data from several areas DNA, patient journals, proteomics, imaging,... Impacts Industry, Environment, Health Societal grand challenges Cheap sequencing technologies results in explosion of DNA data
3 DNA sequencing Reading the order of bases in DNA fragments
4 Why NGS? Transforming how we are doing biological science (and bioinformatics) by allowing experiments that could not have been done before, and perform experiments much faster
5 How? by producing massive amounts of sequence data, really fast
6 First generation: Sanger Fragment DNA Clone into plasmid and amplify Sequence using dntp + labelled ddntps (stops reaction) Run capillary electrophoresis/ gel and read DNA code Low output, long reads (~ nt), high quality
7 1st generation to NGS 1,000,000,000 Single molecule? Illumina Kilobases per day per machine 100,000,000 10,000,000 1,000, , ,000 1, Gel-based systems Manual slab gel Automated slab gel Massively parallel sequencing Capillary sequencing First-generation capillary Microwell pyrosequencing Second-generation capillary sequencer Short-read sequencers Solid 454 Ion Torrent Pacific Biosciences Oxford Complete Nanopore Genomics Future Sanger Chain-termination method Year Human genome 10X Genomics Stratton et al., Nature 2009
8 Sequencing costs Computer speed and storage capacity is doubling every 18 months and this rate is steady DNA sequence data is doubling faster than computer speeds!
9 Human sequencing First draft genome of human in 2001, final 2004 Estimated costs $3 billion, time 13 years Today: $ for one genome A couple of days!
10 Storage and analysis Highest cost is (almost) not the sequencing but storage and analysis A standard human (30-40x) wholegenome sequencing exp. would create 150 Gb of data One Illumina HiSeq X system: 18,000 human genomes per year!
11 Distributed data production World wide >900 centers >60 Pb pr year (2014) Data transfer and storage becomes an issue
12 The X Genomes projects Human population projects 1000 genomes project (2500 individuals) Genomics England (100k individuals) US Precisions medicine (1 million individuals) 100K pathogens project, Earth Microbiome project, Cancer genome project, Plants and animals, Insects,
13 NGS in the clinic Diagnostics of patients (+ fetus) Focused treatment of cancer patients Sequencing of bacterial isolates Country-wide projects: UK, US, Qatar, Finland, China, DK: Danish regions want to sequence 100k individuals Ethical debate?
14 NGS & Bioinformatics Extreme data size causes problems Just transferring and storing the data Standard comparisons fail (N*N) Standard/old tools can not be used Think in fast and parallel programs
15 What can we use it for? Whole genome re-sequencing Population genomics Diagnostics Cancer genomics Ancient genomes Metagenomics RNA sequencing Chip-seq Genomic Epidemiology anything with DNA
16 How it works?
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